Fast free of acrylamide clearing tissue (FACT) for clearing, immunolabelling and three-dimensional imaging of partridge tissues

Fast free of acrylamide clearing tissue (FACT) is a modified sodium dodecyl sulfate-based clearing protocol for the chemical clearing of lipids that completely preserves fluorescent signals in the cleared tissues. The FACT protocol was optimized to image translucent immunostained brain and non-nervous tissues. For this purpose adult male Chukar partridge (Alectoris chukar) was used as a model. After clearing the tissues, 1 or 2 mm-thickness sections of tissues were immunolabeled. The paraventricular nucleus in the hypothalamus (2-mm section) was cleared with FACT, and then was stained with gonadotropin-inhibitory hormone (GnIH) antibody and Hoechst. Simultaneously, immunohistochemical (IHC) staining of cryosectioned brain (30 μm) was done by GnIH-antibody. The FACT protocol and staining of cell nuclei of nine other tissues were done by a z-stack motorized fluorescent microscope. GnIH-immunoreactive neurons were found by FACT and IHC during the breeding season in male partridges. Deep imaging of the kidney, duodenum, jejunum, lung, pancreas, esophagus, skeletal muscle, trachea, and testis were also done. The FACT protocol can be used for the three-dimensional imaging of various tissues and immunostained evaluation of protein markers

Formalin Effects on the Nose and Throat of Personnel of Anatomical Sciences Departments in Iran Medical Schools

Abstract Background: Formaldehyde (and its compositions) is a chemical substance that is widely used in chemical industries (detergents, cosmetics and so on), medicine (for sterilization of operation room and its instruments and fixation of histological specimens and cadavers) and even in textiles and papers. Methods: In recent years many studies have been done in formalin effects on people health. But anybody did not study formalin effects on nose and throat in persons that have direct contact with this substance in Iran. For this reason; questionnaires were sent to personnel of anatomical sciences departments in medical schools were selected as experimental group and personnel of high schools as control group. Questionnaires were prepared that had following indexes: Nasal signs (runny nose and smell sense changes), throat signs (sore throat), occupation, sex, age, formalin contact (duration and direct or indirect contact) and diseases history. Data were analyzed by Chi-square test. Results: Results showed that contact with formalin causes decrease (or loss) of smell sense and increase of runny nose and sore throat. These changes have direct relation with contact duration and direct contact. Relation between contact with formalin and these changes is significant (P Conclusions: Formalin decreases smell sense and increases runny nose and sore throat. It is suggested that people should not have any direct contact with formalin. Also, departments of anatomical sciences use low formalin (or formalin free) agents for fixation of cadavers

Optimisation of ketamine‐xylazine anaesthetic dose and its association with changes in the dendritic spine of CA1 hippocampus in the young and old male and female Wistar rats

Abstract Background A combination of ketamine‐xylazine (K‐X) is frequently used for anaesthesia in rats. Sex and age affect this cocktail dosage. Ketamine causes a hypnotic effect by blocking NMDA receptors located on the dendritic spine of the CA1 region. Objectives The present study aimed to find the optimal dosage of K‐X and its association with the changes in dendritic spine number of the CA1 region for aged and young rats of both sexes. Methods We injected 150–4 mg/kg of K‐X in young and 100–2 mg/kg in aged Wistar rats intraperitoneally and recorded the onset time and duration of anaesthesia and death percentage. Then, animals were sacrificed, brains removed, cut and after Golgi‐Cox staining, the total number of dendritic spines on CA1 was estimated. Results The findings showed that the onset time of anaesthesia lasted longer and its duration lasted shorter, and the number of mature spines decreased with aging, but sex caused no significant effect. The death percentages in young groups comprise 20% and in the aged groups were lower: 5% in males and 0.0% in females. Conclusions It seems 100–2 mg/kg of K‐X is an optimal dose in aged rats and retains an association with reduction of the mature dendritic spine of CA1

Therapeutic Effects of Dimethyl Fumarate on the Rat Model of Brain Ischemia

Blood-brain barrier (BBB) disruption, inflammation, and cell death are major pathogenic mechanisms in ischemic stroke. Dimethyl fumarate (DMF) has anti-inflammatory and immune-modulatory effects. So, this study aimed to elucidate the effects of DMF on brain ischemia in the middle cerebral artery occlusion (MCAO) model. 69 Sprague-Dawley male rats were allocated into a sham group that was just subjected to surgery stress; vehicle and DMF groups, after MCAO, received vehicle or 30 mg/kg DMF for three days. Neurological scores were evaluated every day. BBB disruption was evaluated by the extravasation of Evans blue. In addition to the measurement of brain water content, the total and infarct volume, numerical density, and the total number of neurons, non-neurons, and dead neurons in the right cortex were estimated by stereological methods. RT-PCR was done to analyze the expression levels of NF-κB and Nrf2. Although brain ischemia treatment with DMF did not have a significant effect on the infarction size, it improved neurobehavioral function, BBB disruption, cerebral edema, increased number of neurons, and expression of Nrf2. It also decreased the number of dead neurons and the expression of NF-κB. DMF beneficial effects on stroke may be mediated through both increase of the Nrf2 and decrease of NF-κB expression

The medial prefrontal cortex to the medial amygdala connections may affect the anxiety level in aged rats

Abstract Background Aging changes brain function and behavior differently in male and female individuals. Changes in the medial prefrontal cortex (mPFC)–medial amygdala (MeA) connectivity affect anxiety‐like behavior. Objectives Therefore, this study aimed to investigate the effect of aging and sex on the mPFC–MeA connection and its association with the level of anxiety‐like behavior. Methods We divided the Wistar rats into the male and female young rats (2‐3‐month‐old) and male and female old rats (18–20 months old). First, the open field test (OFT) was performed, and then 80 nl of Fluoro‐Gold (FG) was injected by stereotaxic surgery in the right or left MeA. After 10 days, the animals were perfused, their brain removed, coronal sections cut, and the number of FG‐labeled cells in the right and left mPFC of each sample was estimated. Results Based on our results, old animals revealed less anxiety‐like behavior than young ones, and young females were less anxious than young males, too. Interestingly, MeA of old male rats received more fibers from the bilateral mPFC than young ones. Also, this connection was stronger in the young females than young males. Altogether, the present study indicated that old individuals had less anxiety‐like behavior and stronger mPFC–MeA connection, and young female rats were less anxious and had a stronger connection of mPFC–amygdala than males of the same age. Conclusion Thus, it seems that there is a negative relationship between anxiety levels based on the rat's performance in the OFT apparatus and the mPFC–MeA connection

Curcumin Effects on Memory Impairment and Restoration of Irregular Neuronal Distribution in the Hippocampal CA1 Region After Global Cerebral Ischemia in Male Rats

Introduction: Global Cerebral Ischemia (GCI) causes neuronal damage with subsequent neurological and cognitive impairments. Curcumin has anti-inflammatory, antioxidant, and neuroprotective properties, which makes it a potential candidate for improving GCI-induced impairments. This study aimed to investigate the effects of curcumin on the neurological and memory deficits, as well as spatial neuronal distribution in the CA1 region after GCI in rats. Methods: 56 Sprague-Dawley male rats were randomly assigned into 4 groups of sham (n=14), control (n=14), curcumin 50 mg/kg (n=14), and curcumin 100 mg/kg (n=14). Each group was divided into the two subgroups of short-term (7 days) and long-term (28 days) treatment periods. The neurological severity score (NSS), passive avoidance task, and the traction test were performed at postoperative days of 0, 1, 2, 3, 7, 14, 21, and 28. The novel object recognition test and Voronoi tessellation were carried out on days 7 and 28 after GCI. Results: Curcumin 100 mg/kg significantly decreased neurological severity score on postoperative days of 7 and 28 compared with the control (P<0.001) and curcumin 50 mg/kg groups (P<0.05-P<0.001). Also, curcumin 100 mg/kg significantly increased step-through latency times on postoperative days of 3-28 and 14-28 compared with the control (P<0.05-P<0.001) and curcumin 50 mg/kg groups (P<0.01-P<0.001). Moreover, it increased the novelty preference index during the novel object recognition test in the 28-day treatment subgroup after GCI. Curcumin (100 mg/kg) could maintain the neuronal aggregation in the CA1 region after GCI at a level near to what is generally observed in normal rats. Conclusion: Curcumin could improve memory and neurological deficits and restore irregular neuronal distribution in the CA1 region after GCI in a time-dependent manner, and its higher dose was more effective than its lower dose. Curcumin may have beneficial effects on reducing brain complications after ischemia

Foramen Tympanicum or Foramen of Huschke: A Bioarchaeological Study on Human Skeletons from an Iron Age Cemetery at Tabriz Kabud Mosque Zone

The foramen tympanicum is an anatomical variation that is created in the tympanic plate of temporal bone during the first year of life. The tympanic plate grows and foramen tympanicum is gradually closed by about the fifth postnatal year. However, due to a defect in normal ossification, foramen tympanicum sporadically remains throughout life. The construction of a shopping center in Tabriz, northwest of Iran, led to the discovery of an Iron Age cemetery (1500-500 BC). Several tombs have been uncovered below one meter of sterile soil so far and a thick level of architectural debris from the medieval city has been discovered. Up to now, no bioarchaeological data has been gathered about the burials in this area. Thus, the present study aimed to evaluate the prevalence of foramen tympanicum in this area. In this study, 45 skeletons were studied and the prevalence of this foramen was about 4.4% bilaterally. We also reported on two babies with fused and un-fused squamotympanic fissure. The persistence of this foramen is a possible risk factor for otologic complications after arthroscopy of the temporomandibular joint and salivary gland fistula through this foramen. The closure of this foramen could be also used for age estimation in sub-adult individuals. The incidence of this trait in this study was similar to other available studies on modern skeletons